萘乙酸对燕麦种子胚根长、胚芽长和发芽率的影响

以燕麦种子为试验材料,研究不同浓度萘乙酸（NAA）营养液对燕麦胚根长、胚芽长和发芽率的影响。结果表明：不同浓度的萘乙酸培养液对燕麦的生长有不同的影响,即有不同程度的促进或抑制作用。其中0.001mg/LNAA对胚根的生长表现出促进作用;0.01mg/LNAA对胚芽长生长表现出促进作用;0.0001mg/LNAA对发芽率有促进作用：0.1~10mg/LNAA对燕麦的胚根长、胚芽长和发芽率均有抑制作用。     

无籽西瓜胚轴维管束转位的观察

借助石蜡切片技术对无籽西瓜胚轴进行连续切片和系统镜检。在根茎过渡区连续横切面上,可看到由根部四原型外始式初生木质部与初生韧皮部相间排列的维管束转变到茎部成为四原型、内始式和双韧并生排列的维管束。转化经过分叉、倒转和合并,是不同步的,从左到右沿顺时针力向逐步转变。中胚轴子叶迹发生处延伸出2个小束。根、茎维管束转换区发生在下胚轴。该区外形较粗大,生理活性较强,对外界条件变化较敏感,是瓜苗抗性最弱部位,也是组培择取外植体最合适的节段。可为瓜苗耕作管理和择取外植体提供形态学参考指标。     

马铃薯原生质体游离与培养体系的研究

普通马铃薯四倍体栽培种“甘农薯2号”、“Favorita敗ⅰ癛usset Burbank”试管苗叶片为材料来源,游离培养马铃薯原生质体。结果表明,游离前材料的低温预处理,酶解前对组织和酶液进行真空渗透处理,均能显著提高原生质体产量。用Ficoll密度梯度离心法收集原生质体进行培养,结果表明第三、四层界面的原生质体有较好的培养效果。培养液中加入10 %～15 % 的马铃薯提取液并进行愈伤组织看护培养,有良好的培养效果。     

秦巴蛹虫草原生质体的制备及再生条件研究

【目的】研究秦巴蛹虫草原生质体制备及再生的最适条件,建立高效制备和再生原生质体的方法。【方法】以秦巴蛹虫草无性型菌株CM-16为材料,研究细胞壁裂解酶种类、酶解时间、酶解温度、菌丝体菌龄及稳渗剂种类对原生质体制备及再生效果的影响,并在单因素试验结果基础上进行正交优化试验。【结果】(1)以0.6mol/L NaCl为稳渗剂,用混合酶(质量分数1%纤维素酶和质量分数0.5%蜗牛酶按1∶1体积比混合)在(26±1)℃对菌龄6d的菌丝酶解3h,原生质体的产量最高。(2)以0.6mol/L甘露醇为稳渗剂,用混合酶(质量分数1%纤维素酶和质量分数0.5%蜗牛酶按1∶1体积比混合)在(26±1)℃对菌龄5d的菌丝酶解2h,原生质体的再生率最高;验证试验结果表明,在此条件下,原生质体再生率平均为25.7%,此时原生质体平均产量为83.42×105 mL-1。【结论】对酶解温度、酶解时间、菌丝体菌龄、稳渗剂及酶系种类等条件的优化,可提高秦巴蛹虫草原生质体的产量及再生率。     

小立碗藓细胞重新编程过程中染色体重塑观察

小立碗藓(Physcomitrella patens)是研究植物发育的理想模式植物,也是目前发现的具有高频率同源重组特性的植物,其再生能力很强。此研究以生长7d的小立碗藓原丝体、培养0d的原生质体和再生2d的原生质体为材料,旨在了解小立碗藓细胞重新编程过程中染色体结构的动态变化。透射电镜观察发现原丝体染色体结构浓缩程度高;而当原丝体脱去细胞壁成原生质体状态时,异染色质开始解凝,染色体结构变得较为松散;原生质体再生2d时其染色体松散程度次之。通过进一步荧光定量PCR分析染色体重塑相关基因SWI/SNF的表达水平,发现这些基因在原生质体中表达量最高。     

Diversity and association analysis of important agricultural trait based on miniature inverted-repeat transposable element specific marker in Brassica napus L.

Miniature inverted-repeat transposable elements(MITEs)are a group of DNA transposable element(TE)which preferentially distributed with gene associated regions.Tens of MITEs families have been revealed in Brassica napus genome,they scatter across the genome with tens of thousands copies and produce polymorphisms both intra-and inter-species.Our previous studies revealed a Tourist-like MITE,Monkey King,associated with vernalization requirement of B.napus,however there are still few studies reveal MITE association with agricultural traits in B.napus.In the present study,80 polymorphic markers were developed from 55 MITEs,and used to evaluate genetic diversity in a panel of B.napus accessions consisting of 101 natural and 25 synthetic genotypes.Five agricultural traits,oil content,glucosinolate content,erucic acid content,weight of thousand seeds(WTS)and plant height,were investigated across 3-years field experiments,in addition,two traits,hypocotyl length and root length,were evaluated at the 4-leaf stage in the laboratory.Correlations between the MITE-based markers and seven traits were analyzed,finally,10 polymorphic markers produced by 6 pairs of MITE specific primers were revealed relatively high correlation with 5 traits.Two polymorphic markers were anchored with two candidate genes,BnaA02g13530D and BnaA08g20010D,respectively,which may contribute to glucosinolate content and WTS.This research may contribute to genetic improvement through utilization of MITE-induced polymorphisms in Brassica species.     

五叶地锦离体培养及植株再生

以五叶地锦(Parthenocissus quinquefolia)子叶柄、下胚轴、叶片为外植体进行离体培养,以1/2 MS、B5和Heller为基本培养基,附加不同浓度的细胞分裂素(6-BA,KT, TDZ)及生长素(NAA)诱导下胚轴直接再生不定芽.实验结果表明,暗处理30 d,靠近下胚轴的子叶柄接种在1/2 MS 0.3 mg·L-16-BA培养基上,其不定芽分化率最高达64.67%;下胚轴接种在1/2 MS 2.0 mg·L-1KT 0.05 mg·L-1NAA和Heller 0.5 mg·L-1BA 0.1 mg·L-1NAA培养基上,其不定芽分化率接近,达到24%左右;无菌苗的叶片接种在B5 0.5 mg·L-1TDZ 0.05 mg·L-1NAA培养基上,其不定芽分化率达17.7%.芽增殖培养基为1/2 MS 0.5 mg·L-16-BA 0.1 mg·L-1NAA增殖系数为3～5, 1/2 MS 0.5 mg·L-1IBA 500 mg·L-1活性炭适于再生幼苗的生根,生根苗经移栽全部成活.     

农杆菌介导胡萝卜转化体系的诱导培养基优化

高频再生系统是外源基因成功转化的先决条件,但再生系统不等同于转化系统。为提高农杆菌共培养后胡萝卜的再生频率,以日本新黑田五寸参和林丰改良五寸参2个品种胡萝卜下胚轴为外植体,利用农杆菌介导法将人源白细胞介素-2基因(IL-2)导入其中,探讨了共培养后诱导培养基中激素浓度和筛选剂对再生的影响。结果显示：不同基因型胡萝卜的分化对激素浓度的反应不同,林丰改良五寸参在3种培养基上的胚性愈伤率差异不显著;日本新黑田五寸参胚性愈伤率和每个外植体分化的抗性株数随激素浓度的提高而呈上升趋势,2,4-D和6-BA浓度为1.0mg/L时胚性愈伤率和每个外植体分化的抗性株数最高,分别为87.50%和59株;日本新黑田五寸参以bar基因为标记基因、PPT作筛选剂时,分化较好。因此,遗传转化过程中,为提高共培养后胡萝卜胚状体的分化率,对共培养后的诱导培养基进行再优化是必要的。     

Effects of insect predation on hypocotyl survival and germination success of maturequercus variabilis acorns

The rates of hypocotyl and radicle survival and of germination success were investigated in mature acorns ofQuercus variabilis Blume in relation to endosperm loss due to seed insects. The acorns were damaged by curculio weevils (Coleoptera: Curculionidae) and moths, including tortricid moths (Lepidoptera: Tortricidae); the former were more abundant than the latter. Acorns damaged by curculio weevils showed a significantly lower germination rate when there was a large endosperm loss than the rate for sound acorns. The survival rate of the hypocotyl and radicle also decreased as endosperm loss increased. These results suggest that acorn germination is directly inhibited by damage to the hypocotyl and radicle, which depends on the amount of endosperm eaten by the weevils. However, the germination rate of acorns with hypocotyl and radicle was consistently high, irrespective of the degree of endosperm loss, which suggests that weevil-damaged acorns probably germinate, provided the hypocotyl and radicle survive until the cessation of damage. Research Fellow of the Japan Society for the Promotion of Science. This study was supported in part by Grants-in-Aid for Scientific Research from the Japan Ministry of Education, Science and Culture (No. 09001647, No. 10660144, and No. 11460068) and the Nippon Life Insurance Foundation.     

A Regeneration System for Cotton Variety Xinluzao 45 via Somatic Embryogenesis

[Objective] The purpose of this study was to expand upland cotton genotypes suitable for regeneration via somatic embryogenesis, so as to provide excellent receptor materials for the genetic transformation of cotton. [Method] Six different hormone combinations were studied to establish a somatic embryo regeneration system for hypocotyl explants of Xinluzao 45, a widely planted variety in Xinjiang Province. [Result] Faint yellow callus was induced on a medium consisting of hormone combinations C1 (0.1 mg·L^－1 acetic acid dichlorobenzene oxide [2, 4-D] + 0.1 mg·L^－1 kinetin [KT]) or C5 (0.1 mg·L^－1 2, 4-D + 0.5 mg·L^－1 indo-3-butyric acid + 0.1 mg·L^－1 KT). The generated callus was able to successfully induce embryogenic callus on hormone-free medium containing doubled KNO3. The embryogenic callus then developed into somatic embryos and regenerated plants. Maximum rates of callus induction and regeneration were 92.2% and 40.3%, respectively. Although no significant difference was observed in the rate of callus induction of Xinluzao 45 hypocotyls between hormone combinations C1 and C5, the incidence of induced embryonic callus was higher on C1 than on C5. [Conclusion] The somatic embryo regeneration system developed in this study provides technical support that will facilitate Agrobacterium-mediated transformation of upland cotton Xinluzao 45.